Zhang Yu, Sun Jing, Wei Yongwei, Li Jianrong
Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 311 Goss Laboratory, 1925 Coffey Road, Columbus, OH, 43210, USA.
Methods Mol Biol. 2016;1404:103-121. doi: 10.1007/978-1-4939-3389-1_7.
Avian metapneumovirus (aMPV), also known as avian pneumovirus or turkey rhinotracheitis virus, is the causative agent of turkey rhinotracheitis and is associated with swollen head syndrome in chickens. aMPV belongs to the family Paramyxoviridae which includes many important human pathogens such as human respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and human parainfluenza virus type 3 (PIV3). The family also includes highly lethal emerging pathogens such as Nipah virus and Hendra virus, as well as agriculturally important viruses such as Newcastle disease virus (NDV). For many of these viruses, there is no effective vaccine. Here, we describe a reverse genetics approach to develop live attenuated aMPV vaccines by inhibiting the viral mRNA cap methyltransferase. The viral mRNA cap methyltransferase is an excellent target for the attenuation of paramyxoviruses because it plays essential roles in mRNA stability, efficient viral protein translation and innate immunity. We have described in detail the materials and methods used to generate recombinant aMPVs that lack viral mRNA cap methyltransferase activity. We have also provided methods to evaluate the genetic stability, pathogenesis, and immunogenicity of live aMPV vaccine candidates in turkeys.
禽偏肺病毒（aMPV），也被称为禽肺病毒或火鸡鼻气管炎病毒，是火鸡鼻气管炎的病原体，与鸡的肿头综合征有关。aMPV属于副粘病毒科，该科包括许多重要的人类病原体，如人类呼吸道合胞病毒（RSV）、人类偏肺病毒（hMPV）和人类副流感病毒3型（PIV3）。该科还包括高致死性新兴病原体，如尼帕病毒和亨德拉病毒，以及具有农业重要性的病毒，如新城疫病毒（NDV）。对于这些病毒中的许多病毒，目前尚无有效的疫苗。在此，我们描述了一种通过抑制病毒mRNA帽甲基转移酶来开发减毒活aMPV疫苗的反向遗传学方法。病毒mRNA帽甲基转移酶是副粘病毒减毒的一个极佳靶点，因为它在mRNA稳定性、高效病毒蛋白翻译和固有免疫中发挥着重要作用。我们详细描述了用于产生缺乏病毒mRNA帽甲基转移酶活性的重组aMPV的材料和方法。我们还提供了评估火鸡中减毒活aMPV候选疫苗的遗传稳定性、致病性和免疫原性的方法。
